title: Disruption of the F-actin cytoskeleton and monolayer barrier integrity induced by PAF and the protective effect of ITF on intestinal epithelium.
abstract: To explore whether platelet-activating factor (PAF) can disrupt the intestinal epithelial barrier directly and is associated with structural alterations of the F-actin-based cytoskeleton, and to observe the protective effect of intestinal trefoil factor (ITF), we establish an intestinal epithelia barrier model using Caco-2 cells in vitro. Transepithelial electrical resistance and unidirectional flux of lucifer yellow were measured to evaluate barrier permeability; immunofluorescent staining and flow cytometry were applied to observe morphological alterations and to quantify proteins of the F-actin cytoskeleton: the tight junction marker ZO-1 and Claudin-1 were observed using immunofluorescent staining. PAF significantly increased paracellular permeability, at the same time, F-actin and tight junction proteins were disrupted. It was thought that ITF could reverse the high permeability by restoring normal F-actin, ZO-1 and Claudin-1 structures. These results collectively demonstrated that PAF plays an important role in the regulation of mucosal permeability and the effects of PAF are correlated with structural alterations of the F-actin-based cytoskeleton and of tight junctions. ITF can protect intestinal epithelium against PAF-induced disruption by restricting the rearrangement of the F-actin cytoskeleton and of tight junctions.
authors.affiliation: Department of Pediatrics, Shengjing Hospital of China Medical University, Shenyang, China.
authors.forename: Ling-fen
authors.initials: LF
authors.lastname: Xu
authors.forename: Cheng
authors.initials: C
authors.lastname: Xu
authors.forename: Zhi-Qin
authors.initials: ZQ
authors.lastname: Mao
authors.forename: Xu
authors.initials: X
authors.lastname: Teng
authors.forename: Li
authors.initials: L
authors.lastname: Ma
authors.forename: Mei
authors.initials: M
authors.lastname: Sun
mesh_terms: D000199:Actins / Q000737:chemistry / Q000502:physiology*; D018938:Caco-2 Cells; D002463:Cell Membrane Permeability; D062445:Claudin-1; D003599:Cytoskeleton / Q000502:physiology* / Q000648:ultrastructure; D004305:Dose-Response Relationship, Drug; D005453:Fluorescence; D006801:Humans; D007413:Intestinal Mucosa / Q000187:drug effects / Q000378:metabolism*; D008565:Membrane Proteins / Q000032:analysis; D010455:Peptides / Q000494:pharmacology*; D010972:Platelet Activating Factor / Q000494:pharmacology*; D019108:Tight Junctions / Q000502:physiology* / Q000648:ultrastructure; D000071164:Trefoil Factor-2
chemical_list: D000199:Actins; C568108:CLDN1 protein, human; D062445:Claudin-1; D008565:Membrane Proteins; D010455:Peptides; D010972:Platelet Activating Factor; D000071164:Trefoil Factor-2